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Factor VIII therapy�

Hady Ahmed El-Barhimy

El-Sadat STEM School

Menofia Government

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Contents

  1. What is hemophilia?
  2. Types of hemophilia.
  3. Hemophilia A
  4. Factor VIII
  5. Function of factor VIII
  6. Mechanism
  7. Genetic circuit
  8. Regulation of F8
  9. Expected results

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What is hemophilia?

  • Hemophilia is a disease that results in a decrease in blood clotting factors.

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  • It is inherited through X chromosomes from the parents.

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  • Its symptoms are bleeding for long times after injury and easy bruising.

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There are four types of hemophilia:

Hemophilia A

Hemophilia B

Hemophilia C

Parahemophilia

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Hemophilia A

  • Hemophilia A is a type of hemophilia where the coagulation factor VIII is deficient.

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  • It usually affects males and is inherited as X chromosome-linked recessive trait.

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Factor VIII

  • Factor VIII is an essential blood clotting protein that is produced in the liver sinusoidal cells and endothelial cells.

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  • It is produced by F8 gene, in which any defect will cause hemophilia A.

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Function of factor VIII

  • It binds with another factor called factor IX.

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  • This interaction causes a series of chemical reactions that form a blood clot.

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  • There is factor VIII medication where the protein is injected in bloodstream by slow intravenous injection.

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Types of factor VIII medication

Factor VIII injection can be produced in two ways:

  1. Plasma derived factor concentrates exploits the coagulation factor from the plasma of a healthy person in order to produce freeze-dried products that can be given to the patients by slow intravenous injection.
  2. Recombinant factor concentrates uses genetically modified organisms to produce the coagulation factor VIII and then collects the produced factor which can be given to the body also by slow intravenous injection. �

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Selection of solution

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  • Why do not we use gene delivery to make the factor VIII protein required to treat the disease.

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  • If a plasmid containing F8 gene is inserted, it will enable the target cell to produce the factor VIII protein required.

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Mechanism

As a solution, we have decided to use a gene delivery technique to transfer the gene required to the liver sinusoidal cells.

We will use Type II restriction enzymes to cut the gene responsible for the synthesis of the viral protein.

This will disable the harm that the virus could create to the body.

Then, we will insert a plasmid that contains F8 gene to synthesize factor VIII protein.

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Selection of gene delivery method

  • We decided to use hepatitis A virus as a gene delivery as its target cell is liver cells which are responsible for the production of factor VIII protein.

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  • We chose hepatitis A instead of any other type of hepatitis as it is the least dangerous and most patients can recover from it without treatment.

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  • The hepatitis A viral protein consists of 3 regions P1, which is the structural protein and P2 and P3 which are non-structural proteins associated with the viral replication.

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Inhibition of viral danger

  • The P1 region of the viral protein is the one that causes the harmful effect of the virus so, if we supress its gene from being translated, the harm will be terminated.

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  • So, if we put a restriction enzyme to cut this part of the viral RNA, we will eliminate the risk from the virus, therefore, making the virus safe to inject in the body.

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Genetic circuit �

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Genetic circuit �code

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Regulation of F8

  • If the factor VIII is in excess amount, the patient will be in a risk of getting deep vein thrombosis.

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  • miRNA-30c-5p and miRNA-374b-5p modulate F8 gene expression to control the amount of factor VIII protein in the blood stream.

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  • So, the plasmid that we will insert must contain one of these micro-RNAs to regulate the synthesis of Factor VIII.

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Cost efficiency

  • The cost of production of the treatment can be quite high as the production of the plasmids and using the viruses is quite costly.

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  • So, this could lead to making the treatment very expensive than using factor VIII injections which will be cheaper.

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  • So, we will try to make a viral factory to produce more viruses by reproducing it using liver endothelial cells and making the treatment using them to lower the cost of the treatment.

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Expected results

The hepatitis A capsid will enter to the liver sinusoidal cells for reproduction without any immune response as the virus has lost its harm to the body.

But instead of synthesizing the viral protein, it will express the F8 gene producing factor VIII to treat the patient from hemophilia A.

When the amount of the protein exceeds the required amount of the body, the micro-RNA inserted in the gene will repress the synthesis of factor VIII.

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Citation

Centers for disease control and prevention. (2022) “Treatment of Hemophilia.” CDC, Retrieved from https://www.cdc.gov/ncbddd/hemophilia/treatment.html

Jankowska, K.I., McGill, J.,Pezeshkpoor, B., Oldenburg, J., Sauna, Z.E. & Atreya C.D. (2020) “Further Evidence That MicroRNAs Can Play a Role in Hemophilia A Disease Manifestation: F8 Gene Downregulation by miR-19b-3p and miR-186-5p.” Retrieved from https://doi.org/10.3389/fcell.2020.00669

Soucie, JM., Miller, CH., Dupervil, B. & Le B, Buckne,r TW. “A New Study of Hemophilia Occurrence Finds Many More Cases in the United States.” CDC, Retrieved from www.cdc.gov/ncbddd/hemophilia/features/keyfinding-hemophilia-occurrence-US.html.

Gouw, S., & van den Berg, H. M. (2009). The multifactorial etiology of inhibitor development in hemophilia: Genetics and environment. Seminars in Thrombosis and Hemostasis, 35(08), 723–734. Retrieved from https://doi.org/10.1055/s-0029-1245105

CDC. (2022, August 1). Diagnosis of hemophilia. Centers for Disease Control and Prevention. Retrieved from

https://www.cdc.gov/ncbddd/hemophilia/diagnosis.html

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Thank you